Monteiro-Hara Alessandra C B A, Jadão Adriana S, Mendes Beatriz M J, Rezende Jorge A M, Trevisan Flavio, Mello Ana Paula O A, Vieira Maria Lucia C, Meletti L M M, De S Piedade Sônia M
Laboratório de Biotecnologia Vegetal, Centro de Energia Nuclear na Agricultura, Universidade de São Paulo (CENA/USP), 13400-970 Piracicaba, SP, Brazil.
Departamento de Fitopatologia e Nematologia, Escola Superior de Agricultura 'Luiz de Queiroz', Universidade de São Paulo (ESALQ/USP), 13418-900 Piracicaba, SP, Brazil.
Plant Dis. 2011 Aug;95(8):1021-1025. doi: 10.1094/PDIS-12-10-0873.
We report on the production and evaluation of passionflower transgenic lines for resistance to Cowpea aphid borne mosaic virus (CABMV). Genetic transformation was done using Agrobacterium tumefaciens and transgene integration was confirmed by Southern blot analyses, resulting in nine transgenic lines for 'IAC 275' and three for 'IAC 277'. Transgenic lines were clonally propagated and evaluated for resistance to CABMV. After the third inoculation, under higher inoculum pressure, only propagated plants of the transgenic line T16 remained asymptomatic, indicating a high resistance to infection with CABMV. This transgenic line was self-pollinated and the R generation was evaluated together with the R generation of another resistant transgenic line (T2) identified previously. Plants were inoculated with CABMV by means of viruliferous Myzus nicotianae. All 524 T2R plants became infected, whereas 13 of 279 T16R remained asymptomatic after four successive inoculations. A T16R generation was obtained and plants were inoculated with CABMV mechanically or by aphids. After successive inoculations, 118 of 258 plants were symptomless, suggesting that the resistance to CABMV was maintained in the plant genome as the homozygous condition was achieved. Five selected resistant T16R plants which contained the capsid protein gene are being crossed for further analyses.
我们报告了用于抗豇豆蚜传花叶病毒（CABMV）的西番莲转基因株系的产生和评估。使用根癌农杆菌进行遗传转化，并通过Southern印迹分析确认转基因整合，得到了9个'IAC 275'转基因株系和3个'IAC 277'转基因株系。对转基因株系进行克隆繁殖并评估其对CABMV的抗性。在第三次接种后，在更高的接种压力下，只有转基因株系T16的繁殖植株保持无症状，表明对CABMV感染具有高抗性。该转基因株系进行自花授粉，并将R代与先前鉴定的另一个抗性转基因株系（T2）的R代一起进行评估。通过带毒的烟草蚜接种CABMV。所有524株T2R植株均被感染，而279株T16R植株中有13株在连续四次接种后仍无症状。获得了T16R代，并对植株进行了CABMV机械接种或蚜虫接种。连续接种后，258株植株中有118株无症状，这表明随着纯合状态的实现，对CABMV的抗性在植物基因组中得以维持。正在对5株含有衣壳蛋白基因的选定抗性T16R植株进行杂交，以进行进一步分析。
